HEMATOLOGY 1 [LEC]
Bachelor of Science in Medical Technology
[PART 1] INTRODUCTION TO ERYTHROCYTES AND ANEMIA
o Deficiency of Iron, Deficiency of erythropoietin, Loss of erythroid precursors due
ANEMIA to autoimmune reaction, Infection, and Infiltration of bone marrow granulomas
• Etiology: “anaimia” = without blood (sarcoidosis)
• Decreased in oxygen carrying capacity of the blood
• CLASSIFICATION ACCORDING TO BLOOD LOSS AND HEMOLYSIS
o RBC morphology • Anemia can also develop as a result of acute blood loss or chronic blood loss
o Etiology/disease mechanism • Increased hemolysis results in a shortened RBC lifespan
• MAJOR MECHANISMS
o Blood loss LABORATORY DIAGNOSIS
o Decreased or Ineffective RBC production COMPLETE BLOOD COUNT WITH RBC INDICES
o Increased RBC destruction (inherited or acquired) RBC INDICES
• Anemia: Hgb and O2 to tissue
• Mean Cell Volume/ Mean Corpuscular Volume: indicates average volume of a
• Main cause: Insufficient Hgb and Impaired function of Hgb single erythrocyte in each blood sample
o MCV = Macrocytic
SIGNS AND SYMPTOMS o IDA, Thalassemia, Siderobalstic anemia, Lead poisoning, Chronic inflammation
• COMMON PHYSICAL MANIFESTATIONS • Mean Corpuscular Hemoglobin: indicated the average weight of Hb per erythrocyte
o Pallor, Low Blood Pressure, Slight fever, Edema o Normocytic
• USUAL SYMPTOMS o Aplastic Anemia, Renal Disease, Splenomegaly Infections, Myelopathies
o Easily fatigability, Dyspnea on exertion • Mean Corpuscular Hemoglobin Concentration: indicates the average
• OTHER MANIFESTATIONS concentration of Hb in the erythrocytes
o Vertigo, Faintness, Headache, Palpitations o MCV = Macrocytic
• PHYSICAL EXAMINATION o Megaloblastic, Myelodysplasia, Liver Diseases
o Skin: Petechia
o Eyes: Jaundice
o Mouth: Bleeding
MECHANISMS OF ANEMIA
INEFFECTIVE ERYTHROPOIESIS
• Refers to the production of erythroid precursor cells that are defective precursors
undergo apoptosis within the bone marrow. RETICULOCYTE COUNT
• Conditions: • Tool to assess bone marrow’s ability to increase
o Megaloblastic anemia: deficient DNA synthesis due to vitamin B12 or folate RBC production in response to anemia
deficiency • Reference range:
o Thalassemia: deficient globin chain synthesis o Adult: 0.5 to 1.5%
o Sideroblastic anemia: deficient protoporphyrin synthesis o Newborn: 2-6%
• Hgb → EPO → RBC Production = Ineffective EPO • Methods:
o Red cells are NOT released o Routine Light Microscope Method
o Calibrated Miller Disk Method
INSUFFICIENT EYTHROPOIESIS • Pale blue or deep blue: Retics
• Refers to decrease in the number of erythroid precursors in the BM, resulting in • Blue-green: mature RBC
decreased RBC production and anemia • Example: A total of 150 retics was seen in square
• Problem: hematopoietic stem cells A after counting 500 RBCS in Square B.
• Causes: o Solution:
150 150
o 𝑥 100 𝑥 100
500 𝑥 9 4500
o = 3.33 %
GONZALES, R. | MED221
HEMATOLOGY 1 [LEC]
Bachelor of Science in Medical Technology
NEW METHOD REAGENTS
New Methylene Blue Method New Methylene
Cook, Meyer, & Tureen Method Brilliant Cresyl Blue
Seiverd’s method Brilliant Cresyl Blue/ NMB • NV: approx. 1%
o The RPI is 1 when the Hct is 0.45
o >3.0 = adequate BM response to anemia
OLD METHOD REAGENTS
o < 2.0 = inadequate BM response
Schilling’s Rapid Method Brilliant Cresyl Blue
HCT% MATURATION TIME (DAYS)
Sabin’s Method Neutral Red and Janus Green
40-45 1.0
Seiverd’s method Brilliant Cresyl Blue/ NMB
35-39 1.5
Osgood-Wilhelm Method NMB
25-34 2.0
15-24 2.5
RETICULOCYTE COUNT
• Increased: Considered as the first sign of accelerated erythropoiesis, Observed in
hemolytic anemias, individuals with iron deficiency anemia, thalassemia, sideroblastic
anemia, acute and chronic blood loss
• Decreased: Aplastic anemia
ABSOLUTE RETICULOCYTE COUNT
• Principle: The ARC reflect the actual number of reticulocytes in one liter of blood BONE MARROW EXAMINATION
• NV: 25-7 x 10^9/L (BROWN); 20-115 x 10^9/L (OTHER REF.) not commonly used • Important findings in the bone marrow that could not point to the underlying cause of
in the laboratory. anemia:
o Abnormality cellularity, Lack of iron-on-iron strains of bone marrow (GOLD
CORRECTED RETICULOCYTE COUNT STANDARD for diagnosis of iron deficiency), Presence of granulomata, fibrosis,
infectious agents, and tumor.
• Principle: The percentage of RTC may appear increased because of early
release into the circulation or because of a decrease in the number of mature cells
LABORATORY FINDINGS
in the circulation. The CRC corrects the observed reticulocyte to a normal Hct of 0.45
L/L to allow correction for the degree of patient anemia. • Major laboratory manifestations of anemia:
• Sometimes referred to as Reticulocyte Index (RI) or Hematocrit correction. o Decreased Hgb, Hct, RBC Count
• Additional Information can be acquired from:
RETICULOCYTE PRODUCTION INDEX o RBC indices and histogram, RBC distribution width, and RBC morphology index
• Also known as Shift Correction
MORPHOLOGIC CLASSIFICATION OF ANEMIA BASED ON MEAN CELL
• Provides a further refinement of the
CRC.
VOLUME
• It is a general indicator of the rate of MICROCYTIC ANEMIA
erythrocyte production increase above • Characterized by an MCV of <80 fL with small RBC (6um)
normal in anemias. • Often associated with hypochromia
• Principle: During intense erythropoietic • RBCS with increased central pallor
stress, the maturation time in the bone • Defective Heme synthesis:
marrow may be shortened to as little as 1 o Iron Deficiency
day, allowing the reticulocytes to circulate o Chronic Inflammatory States
longer than usual in the peripheral blood. Cells released early to the peripheral blood o Defective Protoporphyrin Synthesis
are referred to as shift cells and have a polychromatophilic appearance. ▪ Sideroblastic anemia, Lead poisoning
• Defective Globin chain synthesis:
o Thalassemia and Hemoglobin E Disease
GONZALES, R. | MED221
HEMATOLOGY 1 [LEC]
Bachelor of Science in Medical Technology
MACROCYTIC ANEMIA MORPHOLOGIC CLASSIFICATION OF ANEMIA BASED ON
• Characterized by an MCV of > 100 fL with large RBCs (>8um). RETICULOCYTE COUNT
• MEGALOBLASTIC ANEMIA: caused by conditions that impair synthesis of DNA
o Vitamin B12 deficiency, Folate deficiency, Thiamine deficiency, Myelodysplasia,
Erythroleukemia, Some drugs
• NON-MEGALOBLASTIC ANEMIA
o Aplastic anemia/Bone marrow failure, Chronic liver disease, Alcohol
abuse/Alcoholism, Obstructive jaundice, Post Splenectomy, Reticulocytosis,
Hypothyroidism, Myeloproliferative Disease, Drugs
NORMOCYTIC ANEMIA
• MCV between 80-100 fL
• If RETIC count is increased (Hemolytic Anemia)
o Intrinsic: Membrane defects, Hemoglobinopathies, Enzyme deficiencies
o Extrinsic: Immune-mediated, Non-immune RBC injury
• If RETIC count is normal/decreased
o Aplastic anemia, Anemia of Renal disease, Myelophthisic anemia, Infection
(Parvovirus B19), Anemia of Chronic Inflammation
• Classified based on decreased or ineffective Erythropoiesis or Excessive RBC loss.
• Decreased or ineffective RBC production: Low Reticulocytes
• Excessive RBC loss: Increased Reticulocytes
• RDW is used in the conjunction of MCV to identify the problem in morphological
characteristics of RBC.
• RDW Normal: Homogenous; RDW Increased: Heterogenous
GONZALES, R. | MED221
HEMATOLOGY 1 [LEC]
Bachelor of Science in Medical Technology
IRON STUDIES
• Serum iron, Total iron binding capacity, Ferritin, Transferrin receptors, Protoporphyrin,
Tissue iron
• Functional, Transport, Storage
• TIBC: carrying/ load capacity of transferrin
o there is an inverse relationship between TIBC and iron storage.
o Increased TIBC = low iron; Decreased TIBC = high iron
• Ferritin: storage compartment
• Transferrin: carries Fe3+
o When we are testing serum iron, we check the transferrin because it reflects the
transport compartment of iron.
• Protoporphyrin: precursor of heme so if it is increased, it will lead to acquired problem
in heme synthesis
• Tissue iron: if increased, it will lead to iron overload that is fatal because iron is not
commonly found in tissues.
ASSESSMENT OF BODY IRON STATUS
TYPICAL ADULT MALE DIAGNOSTIC USE AND
LABORATORY ASSAY REFERENCE INTERVAL COMPARTMENT
ASSESSED
Serum iron level 50–160 μg/dL Indicator of available
transport iron
Total iron binding 250–400 μg/dL Total amount of iron that
capacity could be transported in the
plasma; indicator indirect
of transferrin level
Transferrin saturation 20%–55% Percentage of transferrin
iron binding sites that are
occupied by iron; indirect
indicator of transport iron
and transferrin level
Serum ferritin level 40–400 ng/mL Indicator of iron stores
Bone marrow or liver Normal iron stores Visual qualitative
biopsy with Prussian visualized assessment of tissue iron
blue staining stores
sTfR level 1.15–2.75 mg/L Indicator of functional iron
sTfR/log ferritin index 0.63–1.8 available in cells
RBC zinc <80 μg/dL of RBCs Indicator of functional iron
protoporphyrin level available in RBCs
Hemoglobin content of 27–34 pg/cell Indicator of functional iron
reticulocytes available in developing
RBCs
SUPPLEMENTARY TEST OF ANEMIA
• Bone marrow examination, Hgb F concentration, Malarial smear, Platelet count,
Reticulocyte count, G6PD testing, Sickling test, Hgb electrophoresis, Iron studies,
Haptoglobin, LDH activity, Vitamin B12 & Folate, Bilirubin, Antibody Screening and ID,
Direct AHG, Urobilinogen, FOBT
GONZALES, R. | MED221
GONZALES, R. | MED221
